Anti-t cell immunotoxin fusion protein and its therapeutic use

ABSTRACT

A combination comprising (a) an anti-T cell immunotoxin fusion protein comprising (i) a diphtheria or  Pseudomonas  toxin moiety and (ii) a targeting moiety suitable for targeting the fusion protein to T cells and (b) at least one chemotherapeutic agent.

The present invention relates to a method of treating proliferativediseases, in particular leukemias or lymphomas. The invention alsorelates to a combination, combined preparation, pharmaceuticalcomposition and commercial package useful in such a method.

The nature of malignant proliferative diseases like solid tumor diseasesand non-solid tumor diseases is multifactorial. This presents a numberof problems in providing suitable therapies which are both effective andsafe. In particular, it is difficult to predict the effects of combiningtwo or more therapeutic agents. Accordingly, there is a need for newagents and methods suitable for treating malignant proliferativediseases.

The invention provides a method of treating a malignant proliferativedisease in a subject in need thereof comprising administering to thesubject a combination which comprises (a) an anti-T cell immunotoxinfusion protein comprising a diptheria or Pseudomonas toxin moiety and atargeting moiety, and (b) at least one chemotherapeutic agent; acombination comprising (a) and (b) as defined above and optionally atleast one pharmaceutically acceptable carrier, e.g. for simultaneous,separate or sequential use, in particular for the delay of progressionor treatment of a malignant proliferative disease, especially a solid ornon-solid tumor disease, such as leukemia or lymphoma; a pharmaceuticalcomposition comprising such a combination; the use of such a combinationfor the preparation of a medicament for the delay of progression ortreatment of a malignant proliferative disease; and a commercial packageor product comprising such a combination.

More particularly, the present invention provides:

1. A combination comprising (a) an anti T cell immunotoxin fusionprotein comprising (i) a diptheria or Pseudomonas toxin moiety and (ii)a targeting moiety suitable for targeting the fusion protein to T cells;and (b) at least one chemotherapeutic agent; components a) and b) may beadministered simultaneously, separately or sequentially.

2. A pharmaceutical composition comprising (a) an anti-T cellimmunotoxin fusion protein comprising (i) a diphtheria or Pseudomonastoxin moiety and (ii) a targeting moiety suitable for targeting thefusion protein to T cells; and (b) at least one chemotherapeutic agent.

3. A commercial package, e.g. a kit comprising (a) an anti-T cellimmunotoxin fusion protein comprising (I) a diptheria or Pseudomonastoxin moiety and (ii) a targeting moiety suitable for targeting thefusion protein to T cells; and (b) at least one chematherapeutic agent;together with instructions for simultaneous, separate or sequential usethereof. In a method according to the invention.

4. Use of a combination or a pharmaceutical composition as defined abovefor delaying the progression of or for the treatment of a malignantproliferative disease.

5. Use of (a) an anti-T cell immunotoxin fusion protein comprising (i) adiptheria or Pseudomonas toxin moiety and (ii) a targeting moietysuitable for targeting the fusion protein to T cells; and (b) at leastone chemotherapeutic agent; for the preparation of a combination orpharmaceutical composition for use in delaying the progression of or forthe treatment of a malignant proliferative disease.

6. A method for delaying the progression of a malignant proliferativedisease in a subject comprising administering to the subject acombination according to the invention.

A combination which comprises (a) an anti-T cell immunotoxin fusionprotein comprising (i) a diptheria or Pseudomonas toxin moiety and (ii)a targeting moiety suitable for targeting the fusion protein to T cellsand (b) at least one chemotherapeutic agent will be referred tohereinafter as a COMBINATION OF THE INVENTION.

According to experimental finding, in vivo the administration of aCOMBINATION OF THE INVENTION, especially comprising an antineoplasticagent as combination partner (b), results in a beneficial effect, e.g. asynergistic therapeutic effect, e.g. with regard to slowing down,arresting or reversing the neoplasm formation or a longer duration oftumor response, or in other beneficial effects, e.g. less side-effects,an improved quality of life and a decreased mortality and morbidity,compared to a monotherapy applying only one of the pharmaceuticallyactive ingredients used in the COMBINATION OF THE INVENTION, e.g. in thetreatment of a tumor that is refractory to other chemotherapeutics knownas anti-cancer agents. In particular, an increased up-take of thecombination partner (b) in tumor tissue and tumor cells may be observed,when applied in combination with combination partner (a).

A further benefit is that lower doses of the active ingredients of theCOMBINATION OF THE INVENTION can be used, for example, that the dosagesneed not only often be smaller but are also applied less frequently, orcan be used in order to diminish the incidence of side-effects. This isin accordance with the desires and requirements of the patients to betreated.

In the combination, pharmaceutical composition or commercial package ofthe present invention, each of component a) and b), independently, maybe present in free form or in the form of a pharmaceutically acceptablesalt.

The term “combination”, as used herein defines especially a “kit ofparts” in the sense that the components (a) and (b) as defined above canbe dosed independently or by use of different fixed combinations withdistinguished amounts of the combination partners (a) and (b), i.e.,simultaneously or at different time points. The parts of the kit ofparts can then, e.g., be administered simultaneously or chronologicallystaggered, that is at different time points and with equal or differenttime intervals for any part of the kit of parts. Very preferably, thetime intervals are chosen such that the effect on the treated disease inthe combined use of the parts is larger than the effect which would beobtained by use of only any one of the combination partners (a) and (b).The ratio of the total amounts of component (a) to component (b) to beadministered in the combined preparation can be varied, e.g. in order tocope with the needs of a patient sub-population to be treated or theneeds of the single patient which different needs can be due to theparticular disease, age, sex, body weight, etc. of the patients.Preferably, there is at least one beneficial effect, e.g., a mutualenhancing of the effect of the combination partners (a) and (b), e.g. amore than additive effect, additional advantageous effects, less sideeffects, a combined therapeutical effect in a non-effective dosage ofone or both of the combination partners (a) and (b).

The term “delay of progression” as used herein means administration ofthe combination to patients to delay tumor growth or invasiveness and/orformation, development, growth and/or spread of metastases ormicrometastases. Patients may also be in a pre-stage or in an earlyphase of the disease. Depending on the tumor type and the particularcombination used a decrease of the tumor volume can be obtained.

The term “solid tumor” especially means breast cancer, cancer of thecolon and generally the GI tract, respiratory tract tumors, e.g. lungcancer, in particular small-cell lung cancer, and non-small-cell lungcancer, head and neck cancer, brain or other central nervous systemtumors, oral cavity tumors, skeletal system tumors, genitourinarycancer, e.g. cervical, uterine, ovarian, testicles, prostate or bladdercancer; skin tumors, e.g. Kaposi's sarcoma. Non-solid tumors includee.g. tumors of blood or lymphatic system, e.g. leukemias and lymphomas,in particular, T cell leukemias and T cell lymphomas, Hodgkin's disease,non-Hodgkin's lymphoma, Burkitt's lymphoma, AIDS-related lymphomas,multiple myeloma, lymphoid leukemia, myeloid leukemia, acute or chroniclymphocytic leukemia, and other unspecified malignant neoplasms oflymphoid.

By “targeting moiety” it is meant any moiety which is suitable fortargeting the fusion protein to T cells. Thus the targeting moiety ispreferably a group, such as a polypeptide sequence, which recognises orbinds to a molecule present on the surface of T cells. More preferablythe targeting moiety binds specifically or selectively to T cells. Insuch an embodiment, the targeting moiety preferably binds to a moleculewhich is selectively expressed on the surface of T cells. Mostpreferably, the targeting moiety comprises an affinant which selectivelybinds to CD3, such is as an anti-CD3 antibody, particularly as disclosedin WO 01/87982 and WO 00/41474.

Component (a) as used herein includes, but is not limited to the anti-Tcell immunotoxin fusion proteins comprising an immunotoxin, morespecifically a diphtheria toxin moiety, and a targeting moiety, morespecifically a recombinantly produced anti-CD3 antibody moiety (eithermonovalent divalent or multivalent). Component (a) also includes arecombinantly produced diphtheria toxin or other mutant diphtheria toxinmoiety as described in WO 01/87982 or US patent application Ser. No.09/573,797, more specifically an immunotoxin as stated above having arecombinantly produced anti-CD3 moiety from the antibody UCHT1 aantibody, more particularly an immunotoxin having the SEQ ID NOs: 1, 2,3, 4, and 5 even more specifically DT389-sFv(UCHT1) (SEQ ID NO:6) and(Ala)dmDT390-bisFv(UCHT1*) (SEQ ID NO:7) that can be prepared andadministered as described in WO 01/87982 or US patent application Ser.No. 09/573,797, the contents thereof being incorporated herein byreference.

Furthermore, component (a) as used herein includes, but is not limitedto the anti-T cell immunotoxin fusion proteins comprising animmunotoxin, more specifically a pseudomonas toxin moiety, and atargeting moiety, more specifically a recombinantly produced anti-CD3antibody moiety (either monovalent, divalent or multivalent). Thecombination partner (a) also includes a recombinantly producedpseudomonas toxin or other mutant pseudomonas toxin moiety as describedin WO 00/41474, more specifically a pseudomonias immunotoxin as statedabove having a recombinantly produced anti-CD3 moiety from the antibodyUCHT1 antibody, more particularly the immunotoxin scFv(UCHT-1)-PE38 thatcan be prepared and administered as described in WO 00/41474, thecontents thereof being incorporated herein by reference.

The pseudomonas toxin moiety is preferably derived from Pseudomonasexotoxin-A (PE) secreted by Pseudomonas aeruginosa. More preferably thepseudomonas toxin moiety comprises PE38, which is a 38 kDa fragment ofPE lacking Domain 1 a (amino acid residues 1 to 250), and also lackingamino acid residues 365 to 380 numbered from the N-terminal of themature PE polypeptide. In other words PE-38 comprises residues 251 to364 joined to residues 381 to 613 of the mature PE polypeptide.Alternative pseudomonas toxin moieties are described on page 12 of WO00/41474 and in U.S. Pat. No. 5,458,878, incorporated herein byreference.

In an alternative embodiment, the targeting moiety comprisesinterleukin-2 (IL-2) or at least a portion of the binding domainthereof, such that the fusion protein is targeted to T cells expressingthe IL-2 receptor and preferably to T cells expressing the high affinityform of the IL-2 receptor. Suitable targeting moieties comprising IL-2or fragments thereof are disclosed in WO 91/13090 and EP 0517829, thecontents of which are incorporated herein by reference. In a preferredembodiment, the fusion protein comprises a diptheria toxin moiety andIL-2 or a fragment thereof, e.g. amino acid residues 2-133 of IL-2, suchas DAB₄₈₆-IL-2 or DAB₃₈₉-IL-2 as described in EP 0517829.

Suitable chemotherapeutic agents as component b) include e.g.antineoplastic agents or agents used as adjuvants in cancer therapy. Theterm “antineoplastic agents” as used herein includes, but is not limitedto aromatase inhibitors, antiestrogens, topoisomerase I inhibitors,topoisomerase 11 inhibitors, microtubule active agents, alkylatingagents, antineoplastic antimetabolites, platin compounds, compoundsdecreasing the protein kinase activity, in particular non-receptortyrosine kinase and further anti-anglogenic compounds, gonadorelinagonists, anti-androgens, bisphosphonates, trastuzumab and miscellaneousanti-cancer agents, e.g. 6-thioguanidine, hydroxyurea, procarbazine orbleomycin.

The term “aromatase inhibitors” as used herein relates to compoundswhich inhibit the estrogen production, i.e. the conversion of thesubstrates androstenedlone and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to steroids,especially exemestane and formestane and, in particular, non-steroids,especially aminoglutethimide, vorozole, fadrozole, anastrozole and, veryespecially, letrozole. Exemestane can be administered, e.g., in the formas it is marketed, e.g. under the trademark AROMASIN™. Formestane can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark LENTARON™. Fadrozole can be administered, e.g., in the form asit is marketed, e.g. under the trademark AFEMA™. Anastrozole can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark ARIMIDEX™. Letrozole can be administered, e.g., in the form asit is marketed, e.g. under the trademark FEMARA™ or FEMAR™.Aminoglutethimide can be administered, e.g., in the form as it ismarketed, e.g. under the trademark ORIMETEN™.

The term “antiestrogens” as used herein relates to compounds whichantagonize the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen can be administered, e.g., inthe form as it is marketed, e.g. under the trademark NOLVADEX™.Raloxifene hydrochloride can be administered, e.g., in the form as it ismarketed, e.g. under the trademark EVISTA™. Fulvestrant can beformulated as disclosed in U.S. Pat. No. 4,659,516 or it can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark FASLODEX™.

The term “topoisomerase I inhibitors” as used herein includes, but isnot limited to topotecan, irinotecan, 9-nitrocamptothecin and themacromolecular camptothecin conjugate PNU-166148 (compound A1 inWO99/17804). Irinotecan can be administered, e.g., in the form as it ismarketed, e.g. under the trademark CAMPTOSAR™. Topotecan can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark HYCAMTIN™.

The term “topoisomerase II inhibitors” as used herein includes, but isnot limited to the antracyclines doxorubicin (including liposomalformulation, e.g. CAELYX™), epirubicin, idarubicin and nemorubicin, theanthraquinones mitoxantrone and losoxantrone, and the podophillotoxinesetoposide and teniposide. Etoposide can be administered, e.g., in theform as it is marketed, e.g. under the trademark ETOPOPHOS™. Teniposidecan be administered, e.g., in the form as it is marketed, e.g. under thetrademark VM 26-BRISTOL™. Doxorubicin can be administered, e.g., in theform as it is marketed, e.g. under the trademark ADRIBLASTIN™.Epirubicin can be administered, e.g., in the form as it is marketed,e.g. under the trademark FARMORUBICIN™. Idarubicin can be administered,e.g., in the form as it is marketed, e.g. under the trademark ZAVEDOS™.Mitoxantrone can be administered, e.g., in the form as it is marketed,e.g. under the trademark NOVANTRON™.

The term “microtubule active agents” relates to microtubule stabilizingand microtubule destabilizing agents including, but not limited to thetaxanes paclitaxel and docetaxel, the vinca alkaloids, e.g.,vinblastine, especially vinblastine sulfate, vincristine especiallyvincristine sulfate, and vinorelbine, discodermolide and epothilones.Docetaxel can be administered, e.g., in the form as it is marketed, e.g.under the trademark TAXOTERE™. Vinblastine sulfate can be administered,e.g., in the form as it is marketed, e.g. under the trademark VINBLASTINR.P.™. Vincristine sulfate can be administered, e.g., in the form as itis marketed, e.g. under the trademark FARMISTINT™. Discodermolide can beobtained, e.g., as disclosed in U.S. Pat. No. 5,010,099.

The term “alkylating agents” as used herein includes, but is not limitedto busulfan, cyclophosphamide, ifosfamide and melphalan.Cyclophosphamide can be administered, e.g., in the form as it ismarketed, e.g. under the trademark CYCLOSTIN™. Ifosfamide can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark HOLOXAN™.

The term “antineoplastic antimetabolites” includes, but is not limitedto 5-fluorouracil, cytarabine, fludarabine, capecitabine, gemcitabine,methotrexate and edatrexate. Capecitabine can be administered, e.g., inthe form as it is marketed, e.g. under the trademark XELODA™.Gemcitabine can be administered, e.g., in the form as it is marketed,e.g. under the trademark GEMZARM™.

The term “platin compounds” as used herein includes, but is not limitedto carboplatin, cis-platin and oxaliplatin. Carboplatin can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark CARBOPLAT™. Oxaliplatin can be administered, e.g., In the formas it is marketed, e.g. under the trademark ELOXATIN™.

The term “compounds decreasing the protein kinase activity and furtheranti-angiogenic compounds” as used herein includes, but is not limitedto compounds decreasing the activity of the epidermal growth factor(EGF) of the epidermal growth factor (EGF), the vascular endothelialgrowth factor (VEGF), the platelet derived growth factor (PDGF), theprotein kinase C and anti-angiogenic compounds having another mechanismfor their activity and/or compounds inhibiting signal transduction (e.g.non-receptor type tyrosine kinases, in particular imatinib in salt form.

Compounds which decreases the activity of VEGF are especially compoundswhich inhibit the VEGF receptor tyrosine kinase, compounds which inhibita VEGF receptor and compounds binding to VEGF, and are in particularthose compounds, proteins and monoclonal antibodies generically andspecifically disclosed in WO 98/35958, WO 00/09495, WO 00/27820, WO00/59509, WO 98/11223, WO 00/27819 and EP 0 769 947; those as describedby M. Prewett et al in Cancer Research 59 (1999) 5209-5218, by F. Yuanet al in Proc. Natl. Acad. Sci. USA, vol. 93, pp. 14765-14770, December1996, by Z. Zhu et al in Cancer Res. 58, 1998, 3209-3214, and by J.Mordenti et al in Toxicologic Pathology, Vol. 27, no. 1, pp 14-21, 1999;in WO 00/37502 and WO 94/10202; Angiostatimm, described by M. S.O'Reilly et al, Cell 79, 1994, 315-328; and Endostatin™, described by M.S. O'Reilly et al, Cell 88,1997,277-285;

-   compounds which decrease the activity of the epidermal growth factor    (EGF) are especially compounds which inhibit the EGF receptor    tyrosine kinase, compounds which inhibit the EGF receptor and    compounds binding to EGF, and are in particular those compounds    generically and specifically disclosed in WO 97/02266, EP 0 564 409,    WO 99/03854, EP 0520722, EP 0 566 226, EP 0 787 722, EP 0 837 063,    U.S. Pat. No. 5,747,498, WO 98/10767, WO 97/30034, WO 97/49688, WO    97/38983 and, especially, WO 96/33980;-   compounds which decreases the activity of the protein kinase C are    especially those staurosporine derivatives disclosed in EP 0 296 110    (pharmaceutical preparation described in WO 00/48571) which    compounds are protein kinase C inhibitors.

Further anti-anglogenic compounds are thalidomide (THALOMID), SU5416,and celecoxib (Celebrex).

The term “gonadorelin agonist” as used herein includes, but is notlimited to abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., inthe form as it is marketed, e.g. under the trademark ZOLADEX™.

Abarelix can be formulated, eg. as disclosed in U.S. Pat. No. 5,843,901.

The term “anti-androgens” as used herein includes, but is not limited tobicalutamide (CASODEX™), which can be formulated, e.g. as disclosed inU.S. Pat. No. 4,636,505.

The term “bisphosphonates” as used herein includes, but is not limitedto etridonic acid, clodronic acid, tiludronic acid, pamidronic acid,alendronic acid, ibandronic acid, risedronic acid and zoledronic acid.“Etridonic acid” can be administered, e.g., in the form as it ismarketed, e.g. under the trademark DIDRONEL™. “Clodronic acid” can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark BONEFOS™. “Tiludronic acid” can be administered, e.g., in theform as it is marketed, e.g. under the trademark SKELID™. “Pamidronicacid” can be administered, e.g., in the form as it is marketed, e.g.under the trademark AREDIA™. “Alendronic acid” can be administered,e.g., in the form as it is marketed, e.g. under the trademark FOSAMAX™.“Ibandronic acid” can be administered, e.g., in the form as it ismarketed, e.g. under the trademark BONDRANA™. “Risedronic acid” can beadministered, e.g., in the form as it is marketed, e.g. under thetrademark ACTONEL™. “Zoledronic acid” can be administered, e.g., in theform as it is marketed, e.g. under the trademark ZOMETA™.

“Trastuzumab” can be administered, e.g., in the form as it is marketed,e.g. under the trademark HERCEPTIN™.

In each case where citations of patent applications or scientificpublications are given in particular in the compound claims and thefinal products of the working examples, the subject-matter of the finalproducts, the pharmaceutical preparations and the claims is herebyincorporated into the present application by reference to thispublications. Comprised are likewise the corresponding racemates,enantiomers, diastereoisomers, tautomers as well as the correspondingcrystal modifications where present, e.g. hydrates, solvates andpolymorphs, which are disclosed therein. The compounds used as activeingredients in the combinations disclosed herein can be prepared andadministered as described in the cited documents, respectively.

The structure of the active agents identified by code nos., generic ortrade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g. PatentsInternational (e.g. IMS World Publications). The corresponding contentthereof is hereby incorporated by reference.

It can be shown by established test models and in particular those testmodels described herein that a COMBINATION OF THE INVENTION results in amore effective delay of progression or treatment of a malignantproliferative disease compared to the effects observed with the singlecombination partners. The person skilled in the pertinent art is fullyenabled to select a relevant test model to prove the hereinbefore andhereinafter mentioned therapeutic indications and beneficial effects.The pharmacological activity of a COMBINATION OF THE INVENTION may, forexample, be demonstrated in a clinical study or in a test procedure asessentially described hereinafter.

Suitable clinical studies are, for example, open label non-randomized,dose escalation studies in patients with advanced solid tumors. Suchstudies prove in particular the synergism of the active ingredients ofthe COMBINATIONS OF THE INVENTION. The beneficial effects onproliferative diseases can be determined directly through the results ofthese studies or by changes in the study design which are known as suchto a person skilled in the art. Such studies are, in particular,suitable to compare the effects of a monotherapy using the activeingredients and a COMBINATION OF THE INVENTION. Preferably, component(a) is administered with a fixed dose and the dose of component (b) isescalated until the Maximum Tolerated Dosage is reached. In a preferredembodiment of the study, each patient receives daily doses of component(a). The efficacy of the treatment can be determined in such studies,e.g., after 18 or 24 weeks by radiologic evaluation of the tumors every6 weeks.

Alternatively, a placebo-controlled, double blind study can be used inorder to prove the benefits of the COMBINATION OF THE INVENTIONmentioned herein.

The COMBINATION OF THE INVENTION can also be applied in combination withsurgical intervention, mild prolonged whole body hyperthermia and/orirradiation therapy.

One embodiment of the invention relates to the use of a COMBINATION OFTHE INVENTION for the prevention, delay of progression or treatment ofor for the preparation of a medicament for the prevention, delay ofprogression or treatment of malignant proliferative disease, inparticular leukemias and lymphomas, even more particular T-cellleukemias and T-cell lymphomas. T-cell leukemias and lymphomas includebut are not limited to T-cell prolymphotic leukemia, T-cell granularlymphotic leukemia, aggressive NK cell leukemia, hairy-cell leukemia,nasal and nasal-type NK/T cell lymphoma, mycosis fungoides and Sezarysyndrome, angioimmunoblastic T-cell lymphoma, peripheral T-cell lymphomaunspecified, adult T-cell leukemia/lymphoma (HTLV1+), an a plastic largecell lymphoma, primary cutaneous OD-30 positive T-celllymphoproliferative disorders, cutaneous T-cell lymphoma, subcutaneouspanniculitis like T-cell lymphoma, intestinal T-cell lymphoma(+enteropathy), and hepatosplenic gamma/delta T-cell lymphoma.

It is one objective of this invention to provide a pharmaceuticalcomposition comprising a quantity, which is jointly therapeuticallyeffective against a proliferative disease comprising the COMBINATION OFTHE INVENTION. In this composition, components (a) and (b) can beadministered together, one after the other or separately in one combinedunit dosage form or in two separate unit dosage forms. The unit dosageform may also be a fixed combination.

The pharmaceutical compositions for separate administration ofcomponents (a) and (b) and for the administration in a fixedcombination, i.e. a single galenical compositions comprising at leasttwo components (a) and (b), according to the invention can be preparedin a manner known per se and are those suitable for enteral, such asoral or rectal, or parenteral administration to mammals, including man,comprising a therapeutically effective amount of at least onepharmacologically active combination partner alone or in combinationwith one or more pharmaceutically acceptable carriers, especiallysuitable for enteral or parenteral application.

Novel pharmaceutical composition contain, for example, from about 0.1%to about 99.9%, preferably from about 20% to about 60%, of the activecomponents. Pharmaceutical preparations for the combination therapy forenteral or parenteral administration are, for example, those in unitdosage forms, such as sugar-coated tablets, tablets, capsules orsuppositories, and furthermore ampoules. If not indicated otherwise,these are prepared in a manner known per se, for example by means ofconventional mixing, granulating, sugar-coating, dissolving orlyophilizing processes. It will be appreciated that the unit content ofa combination partner contained in an individual dose of each dosageform need not in itself constitute an effective amount since thenecessary effective amount can be reached by administration of aplurality of dosage units.

In particular, a therapeutically effective amount of each activecompound of the COMBINATION OF THE INVENTION may be administeredsimultaneously or sequentially and in any order, and the components maybe administered separately or as a fixed combination. For example, themethod of the invention may comprise (i) administration of the component(a) in free or pharmaceutically acceptable salt form and (ii)administration of at least one component (b) in free or pharmaceuticallyacceptable salt form, simultaneously or sequentially in any order, injointly therapeutically effective amounts, preferably in synergisticallyeffective amounts, e.g. in daily dosages corresponding to the amountsdescribed herein. The individual active components of the COMBINATION OFTHE INVENTION can be administered separately at different times duringthe course of therapy or concurrently in divided or single combinationforms. Furthermore, the term administering also encompasses the use of apro-drug of a component that converts in vivo to the component as such.The instant invention is therefore to be understood as embracing allsuch regimes of simultaneous or alternating treatment and the term“administering” is to be interpreted accordingly.

The effective dosage of each component employed in the COMBINATION OFTHE INVENTION may vary depending on the particular compound orpharmaceutical composition employed, the mode of administration, thecondition being treated, the severity of the condition being treated.Thus, the dosage regimen of the COMBINATION OF THE INVENTION is selectedin accordance with a variety of factors including the route ofadministration and the renal and hepatic function of the patient. Aphysician or clinician of ordinary skill can readily determine andprescribe the effective amount of the single active ingredients requiredto prevent, counter or arrest the progress of the condition. Optimalprecision in achieving concentration of the active ingredients withinthe range that yields efficacy without toxicity requires a regimen basedon the kinetics of the active ingredients' availability to target sites.

An anti-T cell immunotoxin fusion protein comprising a diphtheria toxinmoiety and a targeting moiety (e.g. (Ala)dmDT390-bisFv(UCHT1*)) ispreferably administered to a human in a dosage in the range of about1-40 μg/day, more preferably 1-30 μg/day and most preferably 1-20 p/day.Unless stated otherwise herein, the compound is preferably administeredfrom one to four times per day over a period of 1 to 10, more preferablyfrom 1 to 5 days. This cycle can be repeated every 1 to 4 weeks,preferably every 3 weeks.

An anti-T cell immunotoxin fusion protein comprising a pseudomonas toxinmoiety and a targeting moiety (e.g. scFv(UCHT-1)-PE38) is preferablyadministered to a human in a dosage in the range of about 1-100 μg/day,more preferably 10-70 μg/day and most preferably 30-50 p/day. Unlessstated otherwise herein, the compound is preferably administered fromone to four times per day over a period of 1 to 10, more preferably from1 to 5 days. This cycle can be repeated every 1 to 4 weeks, preferablyevery 3 weeks.

Non-receptor type tyrosine kinases, in particular imatinib in salt form,is preferably administered to a human in a dosage in the range of about2.5 to 850 mg/day. Unless stated otherwise herein, the compound ispreferably administered from one to four times per day.

Fadrozole may be administered orally to a human in a dosage rangevarying from about 0.5 to about 10 mg/day, preferably from about 1 toabout 2.5 mg/day

Exemestane may be administered orally to a human in a dosage rangevarying from about 5 to about 200 mg/day, preferably from about 10 toabout 25 mg/day, or parenterally from about 50 to 500 mg/day, preferablyfrom about 100 to about 250 mg/day. If the drug shall be administered ina separate pharmaceutical composition, it can be administered in theform disclosed in GB 2,177,700.

Formestane may be administered parenterally to a human in a dosage rangevarying from about 100 to 500 mg/day, preferably from about 250 to about300 mg/day.

Anastrozole may be administered orally to a human in a dosage rangevarying from about 0.25 to 20 mg/day, preferably from about 0.5 to about2.5 mg/day.

Aminogluthemide may be administered to a human in a dosage range varyingfrom about 200 to 500 mg/day.

Tamoxifen citrate may be administered to a human in a dosage rangevarying from about 10 to 40 mg/day.

Vinblastine may be administered to a human in a dosage range varyingfrom about 1.5 to 10 mg/m² day.

Vincristine sulfate may be administered parenterally to a human in adosage range varying from about 0.025 to 0.05 mg/kg body weight week.

Vinorelbine may be administered to a human in a dosage range varyingfrom about 10 to 50 mg/m² day.

Etoposide phosphate may be administered to a human in a dosage rangevarying from about 25 to 115 mg/m² day, e.g. 56.8 or 113.6 mg/m² day.

Teniposide may be administered to a human in a dosage range varying fromabout 75 to 150 mg about every two weeks.

Doxorubicin may be administered to a human in a dosage range varyingfrom about 10 to 100 mg/m² day, e.g. 25 or 50 mg/m² day.

Epirubicin may be administered to a human in a dosage range varying fromabout 10 to 200 mg/m² day.

Idarubicin may be administered to a human in a dosage range varying fromabout 0.5 to 50 mg/m² day.

Mitoxantrone may be administered to a human in a dosage range varyingfrom about 2.5 to 25 mg/m² day.

Paclitaxel may be administered to a human in a dosage range varying fromabout 50 to 300 mg/m² day.

Docetaxel may be administered to a human in a dosage range varying fromabout 25 to 100 mg/m² day.

Cyclophosphamide may be administered to a human in a dosage rangevarying from about 50 to 1500 mg/m² day.

Melphalan may be administered to a human in a dosage range varying fromabout 0.5 to 10 mg/m² day.

5-Fluorouracil may be administered to a human in a dosage range varyingfrom about 50 to 1000 mg/m² day, e.g. 500 mg/m² day.

Capecitabine may be administered to a human in a dosage range varyingfrom about 10 to 1000 mg/m² day.

Gemcitabine hydrochloride may be administered to a human in a dosagerange varying from about 1000 mg/week.

Methotrexate may be administered to a human in a dosage range varyingfrom about 5 to 500 mg/m² day.

Topotecan may be administered to a human in a dosage range varying fromabout 1 to 5 mg/m² day.

Irinotecan may be administered to a human in a dosage range varying fromabout 50 to 350 mg/m² day.

Carboplatin may be administered to a human in a dosage range varyingfrom about 200 to 400 mg/m² about every four weeks.

Cisplatin may be administered to a human in a dosage range varying fromabout 25 to 75 mg/m² about every three weeks.

Oxaliplatin may be administered to a human in a dosage range varyingfrom about 50 to 85 mg/m² every two weeks.

Alendronic acid may be administered to a human in a dosage range varyingfrom about 5 to 10 mg/day.

Clodronic acid may be administered to a human e.g. in a dosage rangevarying from about 750 to 1500 mg/day.

Etridonic acid may be administered to a human in a dosage range varyingfrom about 200 to 400 mg/day.

Ibandronic acid may be administered to a human in a dosage range varyingfrom about 1 to 4 mg every three to four weeks.

Risedronic acid may be administered to a human in a dosage range varyingfrom about 20 to 30 mg/day.

Pamidronic acid may be administered to a human in a dosage range varyingfrom about 15 to 90 mg every three to four weeks.

Tiludronic acid may be administered to a human in a dosage range varyingfrom about 200 to 400 mg/day.

Trastuzumab may be administered to a human in a dosage range varyingfrom about 1 to 4 mg/m² week.

Bicalutamide may be administered to a human in a dosage range varyingfrom about 25 to 50 mg/m² day.

The following examples illustrate the invention described above, withoutintending to limit the scope of the invention in any way. The beneficialeffects of the COMBINATION OF THE INVENTION can also be determined byother test models known as such to the person skilled in the pertinentart.

EXAMPLE 1 (Ala)dmDT390-bisFv(UCHT1*) Alone and in Combination withImatinib, Taxol®, or Doxorubicin in Patients with Cuteanous T cellLymphoma

Patients with cutaneous T cell lymphoma are selected. Cells arehistologically assessed for II-2 receptor determined byImmunohistochemical staining using an anti-CD25 antibody and antibodiesdirected against the p75 component of the II-2 receptor (Foss F M etal., Curr Top Microbiol Immunol. 1998; 234: p. 63-81). Only cellshistologically and immunohistochemically representative of malignantcells are assessed for 11-2R. When more than 25% of the tumor cellsstained with either p55 or p75, (Ala)dmDT390-bisFv(UCHT1* treatment isbegun at 9 or 18 μg/kg, i.v. over 5 min daily for 5 days and repeatedevery 3 weeks for a maximal period of 6 cycles. The combination partnerImatinib is administered at 100 mg/kg, p.o., every 12 h, or thecombination partner Taxol® is administered at 15 mg/kg, i.v., every 48hours, for five times, or the combination partner doxorubicin at 9mg/kg, i.v., every 7 days.

Data points just spanning the IC50 of the agents alone or in combinationare entered into the CalcuSyn program (CalcuSyn, Biosoft, Cambridge UK).This program calculates a non-exclusive combination index (Cl), whosevalue is indicative of the interaction of the two compounds, where Cl ˜1represents nearly additive effects; 0.85-0.9 Indicates a slightsynergism and a value below 0.85 indicates synergy.

The study indicates that combination treatment of(Ala)dmDT390-bisFv(UCHT1*) with any of the combination partner has abeneficial effect in preventing and treating cuteanous T cell lymphoma.

EXAMPLE 2 scFv(UCHT-1)-PE38 Alone and in Combination with Imatinib,Taxol®, or Doxorubicin in Patients with Hairy-Cell Leukemia.

Patients with hairy-cell leukemia are selected. All patients havecirculating malignant cells that express CD22, adequate organ function,and an absence of high levels of neutralizing antibodies againstscFv(UCHT-1)-PE38.

Between 0.2 and 4.0, mg of scFv(UCHT-1)-PE38 are diluted in 50 ml of 0.2percent albumin in 0.9 percent sodium chloride and administered as a30-minute intravenous Infusion every other day for a total of threedoses. The combination partner Imatinib is administered at 100 mg/kg,p.o., every 12 h, or the combination patner Taxol® is administered at 15mg/kg, i.v., every 48 hours, for five times, or the combination partnerdoxorubicin at 9 mg/kg, i.v., every 7 days. The disease is assessed byfluorescence-activated cell-sorter (FACS) analysis to detect thehairy-cell leukemia antigen CD22 before and after treatment.

Data points just spanning the IC50 of the agents alone or in combinationare entered into the CalcuSyn program (CalcuSyn, Biosoft, Cambridge UK).This program calculates a non-exclusive combination index (CI), whosevalue is indicative of the interaction of the two compounds, where Cl ˜1represents nearly additive effects; 0.85-0.9 indicates a slightsynergism and a value below 0.85 indicates synergy.

The study indicates that combination treatment of scFv(UCHT-1)-PE38 withany of the combination partner has a beneficial effect in preventing andtreating cuteanous T cell lymphoma.

1. A combination comprising: (a) an anti-T cell immunotoxin fusionprotein comprising (i) a diphtheria or Pseudomonas toxin moiety and (ii)a targeting moiety suitable for targeting the fusion protein to T cells;and (b) at least one chemotherapeutic agent comprising: (i) anantineoplastic agent selected from the group consisting of: aromataseinhibitors, antiestrogens, topoisomerase I inhibitors, topoisomerase 11inhibitors, microtubule active agents, busulfan, ifosfamide, melphalan,5-fluorouracil, cytarabine, fludarabine, capecitabine, gemcitabine,edatrexate, platin compounds, compounds decreasing the protein kinaseactivity and further anti-angiogenic compounds, gonadorelin agonists,anti-androgens, bisphosphonates, trastuzumab, 6-thioguanidine,hydroxyurea, procarbazine and bleomycin; or (ii) an agent used as anadjuvant in cancer therapy.
 2. A pharmaceutical composition comprising:(a) an anti-T cell immunotoxin fusion protein comprising (i) adiphtheria or Pseudomonas toxin moiety and (ii) a targeting moietysuitable for targeting the fusion protein to T cells; and (b) at leastone chemotherapeutic agent comprising: (i) an antineoplastic agentselected from the group consisting of: aromatase inhibitors,antiestrogens, topoisomerase I inhibitors, topoisomerase 11 inhibitors,microtubule active agents, busulfan, ifosfamide, melphalan,5-fluorouracil, cytarabine, fludarabine, capecitabine, gemcitabine,edatrexate, platin compounds, compounds decreasing the protein kinaseactivity and further anti-angiogenic compounds, gonadorelin agonists,anti-androgens, bisphosphonates, trastuzumab, 6-thioguanidine,hydroxyurea, procarbazine and bleomycin; or (ii) an agent used as anadjuvant in cancer therapy.
 3. A combination according to claim 1,wherein the targeting moiety comprises an anti-CD3 antibody.
 4. Acombination according to claim 1, wherein the targeting moiety comprisesIL-2 or at least a portion of the binding domain thereof.
 5. Acombination according to claim 1, wherein the anti-T cell immunotoxinfusion protein comprises a polypeptide having a sequence as defined inany one of SEQ ID NOs 1 to
 7. 6-9. (canceled)
 10. A method of treating amalignant proliferative disease in a subject in need thereof comprisingadministering to the subject a combination as defined in claim
 1. 11. Amethod according to claim 10, wherein the malignant proliferativedisease comprises a lymphoma or leukemia.
 12. A method of treating amalignant proliferative disease in a subject in need thereof comprisingadministering to the subject a pharmaceutical composition as defined inclaim
 2. 13. A method according to claim 12, wherein the malignantproliferative disease comprises a lymphoma or leukemia.
 14. Apharmaceutical composition according to claim 2, wherein the targetingmoiety comprises an anti-CD3 antibody.
 15. A pharmaceutical compositionaccording to claim 2, wherein the targeting moiety comprises IL-2 or atleast a portion of the binding domain thereof.
 16. A pharmaceuticalcomposition according to claim 2, wherein the anti-T cell immunotoxinfusion protein comprises a polypeptide having a sequence as defined inany one of SEQ ID NOs 1 to
 7. 17. A combination according to claim 3,wherein the anti-T cell immunotoxin fusion protein comprises apolypeptide having a sequence as defined in any one of SEQ ID NOs 1 to7.
 18. A combination according to claim 4, wherein the anti-T cellimmunotoxin fusion protein comprises a polypeptide having a sequence asdefined in any one of SEQ ID NOs 1 to
 7. 19. A pharmaceuticalcomposition according to claim 14, wherein the anti-T cell immunotoxinfusion protein comprises a polypeptide having a sequence as defined inany one of SEQ ID NOs 1 to
 7. 20. A pharmaceutical composition accordingto claim 15, wherein the anti-T cell immunotoxin fusion proteincomprises a polypeptide having a sequence as defined in any one of SEQID NOs 1 to 7.